Recombinant Human His6-USP12/UAF1 Complex Protein, CF
Recombinant Human His6-USP12/UAF1 Complex Protein, CF Summary
Product Specifications
Recombinant Human USP12/UAF1 Complex is a Ubiquitin-specific deconjugating enzyme heterodimer. Reaction conditions will need to be optimized for each specific application. We recommend an initial USP12/UAF1 Complex concentration of 50-500 nM when using Ubiquitin-AMC or Ubiquitin-Rhodamine (U-550, U-555) as a substrate. In vitro, the USP12/UAF1 complex will not efficiently hydrolyze poly-Ubiquitin chains in the absence of WDR20.
Contains a C-terminal 6-His tag
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
E-580
| Formulation | X mg/ml (X μM) in 50 mM HEPES pH 8.0, 150 mM NaCl, 10% (v/v) Glycerol, 2 mM TCEP |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: USP12/UAF1 Complex
Ubiquitin carboxyl-terminal hydrolase 12 (USP12) is a specialized cysteine protease with a predicted molecular weight of 43 kDa. USP12 is a member of the peptidase C19 family and USP12/USP46 subfamily, and the human protein shares 98% amino acid sequence identity with its mouse ortholog. Like USP1 and USP46, USP12 lacks appreciable activity in the absence of its co-activator UAF1. Addition of UAF1 increases the in vitro activity of USP12 20-fold or more. WDR20 is often found in complex with both USP12 and UAF1, and the ternary complex is reported to be more active than the USP12/UAF1 complex. Biologically, USP12 has been implicated as a positive regulator of Androgen Receptor (AR) activity via a mechanism that involves down-regulation of Akt activity. As such, it has been proposed that USP12 inhibition could offer a therapeutic target for some forms of prostate cancer. Recombinant USP12 contains a C-terminal 6-His tag. Recombinant UAF1 also contains a C-terminal 6-His tag. The assembled complex contains 25 µg USP12 and 45 µg UAF1, resulting in a 1:1 stoichiometry.
- Burska, U.L. et al. (2013) J Biol Chem. 288: 32641
- Cohn, M.A. et al. (2009) J Biol Chem. 284: 5343
- Gangula, N.R. & Maddika, S. (2013) J Biol Chem. 288: 34545
- Kee, Y. et al. (2010) J Biol Chem. 285: 11252
- McClurg, U.L. et al. (2014) Oncotarget 5: 7081
- Ohashi, M. et al. (2015) PLoS Pathog. DOI: 10.1371/journal.ppat.1004822
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